Harnessing polymicrobial interactions in the catheterized urinary tract to identify novel inhibitors of Proteus mirabilis urease activity

利用插管尿路中的多种微生物相互作用来鉴定奇异变形杆菌尿素酶活性的新型抑制剂

• 批准号: 10678389
• 负责人: Lauren Beryl Guterman
• 金额: $ 3.73万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10678389
• 关键词: Acids; Active Sites; Ammonia; Animals; Antibiotics; Bacteremia; Bacteria; Bathing; Biological Availability; Biomass; Bladder; Carbon Dioxide; Catheterization; Catheters; Cells; Cessation of life; Clinical; Clinical Trials; Communicable Diseases; Cytoplasm; Data; Development; Dose; Drainage procedure; Enterococcus faecalis; Enzymes; Extracellular Space; Foundations; Future; Glass; Goals; Histamine; Hospitalization; Human; Imidazole; In Vitro; Incidence; Incubated; Individual; Indwelling Catheter; Infection; Innate Immune Response; Internal Medicine; Investigation; Ions; Klebsiella aerogenes; Knowledge; Laboratories; Length of Stay; Leu-Gly; Measures; Mentors; Messenger RNA; Microbial Biofilms; Modeling; Morbidity - disease rate; Morganella morganii; National Research Service Awards; Nickel; Organism; Pain; Pathogenesis; Patient-Focused Outcomes; Patients; Physicians; Physiological; Preceptorship; Precipitation; Process; Production; Proteins; Proteus mirabilis; Providencia; Publications; Research; Risk; Science; Scientist; Sepsis; Severities; Severity of illness; Staphylococcus aureus; System; Temperature; Testing; Therapeutic; Time; Tissues; Toxic effect; Training; United States Food and Drug Administration; Urea; Urease; Urinary Calculi; Urinary Catheterization; Urinary tract; Urinary tract infection; Urine; Virulence Factors; Water; Work; antimicrobial; catheter associated UTI; drug discovery; efficacy evaluation; efficacy testing; enhancing factor; experimental study; improved; in vitro Model; in vitro activity; inhibitor; insight; instrumentation; interest; medical schools; metabolomics; metalloenzyme; microbial; mortality; mouse model; new therapeutic target; novel; novel therapeutics; pathogen; phenylpyruvate; pre-clinical assessment; prevent; side effect; translational potential; urinary

Project Summary Catheter-associated urinary tract infection (CAUTI) is the leading cause of secondary nosocomial bloodstream infections. Urease-producing organisms such as Proteus mirabilis are common causes of CAUTI and associated with numerous complications. Urease hydrolyzes urea to ammonia, which raises the urine pH and causes precipitation of ions into crystals, ultimately leading to catheter encrustation and blockage and urinary stone formation and increasing the risk of bacteremia, sepsis and death. The only urease inhibitor approved by the Food and Drug Administration (FDA), acetohydroxamic acid (AHA), shows efficacy in preventing urinary stone formation but has severe side effects that limit its clinical use. Therefore, alternative strategies are needed for targeting bacterial urease activity to prevent catheter encrustation and stone formation. In patients with long-term indwelling catheters, CAUTI is often polymicrobial. As demonstrated in our prior publications and preliminary data, we found that common constituents of polymicrobial CAUTI secrete molecules that modulate P. mirabilis urease activity and infection severity. We therefore performed untargeted global metabolomics analysis on cell-free supernatants of urease-modulating bacterial strains and identified 36 candidate urease dampening metabolites. We validated the activity of four compounds: histamine (CAS 51-45- 6), leucylglycine (CAS 686-50-0), phenylpyruvate (CAS 156-06-9) and imidazole lactate (CAS 14403-45-3). Our preliminary data suggests that leucylglycine and imidazole lactate dampen urease activity, at least in part, by acting directly on the urease active site. In contrast, histamine and phenylpyruvate appear to indirectly inhibit urease activity through an uncharacterized mechanism. The goal of this NRSA F30 proposal is to determine the mechanism of action of urease modulation and conduct pre-clinical assessment of the therapeutic potential of these dampening compounds. Aim 1 will determine the mechanism of indirect urease modulation for histamine and phenylpyruvate and explore whether compounds of different mechanisms of action have synergistic effects on urease activity when administered in combination against a panel of urease- producing pathogens. Aim 2 will assess the translational potential of dampening compounds for preventing Foley catheter encrustation, crystalline biofilm formation, and blockage. The experiments outlined in this proposal will provide a framework for harnessing polymicrobial interactions for drug discovery and the development of novel therapeutics to prevent CAUTI associated morbidity and mortality. This work will take place at the Jacobs School of Medicine and Biomedical Sciences in the laboratory of Dr. Chelsie Armbruster, who is an expert in CAUTI and microbial pathogenesis research. The training plan is tailored for my development as a physician-scientist in the field of infectious disease, and includes mentoring by successful physician-scientists and clinical preceptorships in infectious disease and internal medicine.

项目概要 导管相关性尿路感染（CAUTI）是继发性院内血流的主要原因 感染。产生脲酶的微生物，如奇异变形杆菌，是 CAUTI 的常见原因， 与许多并发症有关。尿素酶将尿素水解成氨，从而提高尿液的 pH 值， 导致离子沉淀成晶体，最终导致导管结垢和堵塞以及尿 结石形成并增加菌血症、败血症和死亡的风险。唯一获批的脲酶抑制剂 美国食品和药物管理局 (FDA) 证明，乙酰异羟肟酸 (AHA) 具有预防尿毒症的功效。 形成结石，但具有严重的副作用，限制了其临床应用。因此，替代策略是 需要针对细菌脲酶活性以防止导管结垢和结石形成。在患者中 长期留置导管的 CAUTI 往往是多种微生物的。正如我们之前的出版物所示 和初步数据，我们发现多种微生物 CAUTI 的常见成分分泌的分子 调节奇异果脲酶活性和感染严重程度。因此，我们进行了非针对性的全球 对脲酶调节菌株的无细胞上清液进行代谢组学分析并鉴定出 36 候选脲酶抑制代谢物。我们验证了四种化合物的活性：组胺（CAS 51-45- 6)、亮氨酰甘氨酸 (CAS 686-50-0)、苯丙酮酸 (CAS 156-06-9) 和咪唑乳酸酯 (CAS 14403-45-3)。 我们的初步数据表明，亮氨酰甘氨酸和咪唑乳酸至少部分抑制脲酶活性， 通过直接作用于脲酶活性位点。相反，组胺和苯丙酮酸似乎间接 通过一种未知的机制抑制脲酶活性。 NRSA F30 提案的目标是 确定脲酶调节的作用机制并进行临床前评估 这些阻尼化合物的治疗潜力。目标1将确定间接脲酶的机制 组胺和苯丙酮酸的调节，并探讨不同机制的化合物是否 当与一组脲酶联合施用时，该作用对脲酶活性具有协同作用 产生病原体。目标 2 将评估阻尼化合物的转化潜力，以预防 福利导管结垢、结晶生物膜形成和堵塞。本文概述的实验 该提案将为利用多微生物相互作用进行药物发现和 开发新疗法以预防 CAUTI 相关的发病率和死亡率。这项工作将需要 位于雅各布斯医学与生物医学科学学院 Chelsie Armbruster 博士的实验室， 他是 CAUTI 和微生物发病机制研究的专家。培训计划是为我量身定制的 作为传染病领域的医师科学家的发展，包括成功人士的指导 传染病和内科领域的医师科学家和临床导师。